Abstract
SOX9 (SRY-related HMG box gene 9) is an essential regulator of male sex determination and testis differentiation in many vertebrate species. However, the functional role of <i>Sox9</i> in testis differentiation has not yet been identified in any reptilian species. Herein, <i>Sox9</i> knockdown in the red-eared slider turtle (<i>Trachemys scripta</i>) embryos at a male-producing temperature led to complete male-to-female sex reversal, characterized by the formation of an ovary-like structure, disappearance of male marker AMH, and ectopic expression of ovarian regulator FOXL2, as well as a female distribution pattern of germ cells. Conversely, <i>in-ovo</i> overexpression of <i>Sox9</i> at a female-producing temperature resulted in partial masculinization of putative female embryos, with the co-existence of AMH and FOXL2. Our study provides the first direct evidence that <i>Sox9</i> is indispensable for testicular differentiation in a reptilian species, further confirming the conserved role of <i>Sox9</i> in vertebrate sexual development.
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